CAJ | 학술논문

目的研究硫链丝菌素通过抑制FoxM1表达对髓母细胞瘤Daoy细胞顺铂(cDDP)化疗敏感性的影响.方法体外常规培养髓母细胞瘤Daoy细胞株,CCK8法检测1、1.25、1.5、1.75、2、3、5μmol/L硫链丝菌素,1、2、3、4、5、10、20、40 μmol/L cDDP及1μmol/L硫链丝菌素与1、2、3、4、5、10、20、40 μmol/L cDDP联合处理后对细胞的抑制率,金式公式分析两药间的互相作用；根据分析结果采用1 μmol/L硫链丝菌素、3μmol/L cDDP单独或联合作用Daoy细胞24 h,流式细胞仪检测细胞凋亡的变化,Western blotting检测FoxM1及凋亡相关蛋白bcl-2、bax、caspase3、聚腺苷二磷酸核糖聚合酶(PARP)表达的变化.结果硫链丝菌素可浓度依赖性地抑制髓母细胞瘤Daoy细胞的生长.金式公式分析显示硫链丝菌素与低浓度(1、2、3、4、5μmol/L) cDDP存在明显的协同作用；流式细胞分析仪显示硫链丝菌素单药组、cDDP单药组细胞凋亡率高于对照组,而联合用药组细胞凋亡率高于硫链丝菌素单药组和cDDP单药组,差异有统计学意义(P＜0.05).Western blotting结果显示,与对照组比较,硫链丝菌素单药组和联合用药组FoxM1蛋白表达明显下降,差异有统计学意义(P＜0.05)；与硫链丝菌素单药组、cDDP单药组比较,联合用药组细胞bcl-2蛋白表达明显下降,而bax、caspase3、PARP蛋白表达明显升高.结论硫链丝菌素和cDDP在低浓度下具有明显的协同抗髓母细胞瘤作用,其机制可能与硫链丝菌素抑制FoxM1表达后增强了cDDP诱导凋亡的能力有关.
목적 연구류련사균소통과억제FoxM1표체대수모세포류Daoy세포순박(cDDP)화료민감성적영향.방법 체외상규배양수모세포류Daoy세포주,CCK8법검측1、1.25、1.5、1.75、2、3、5μmol/L류련사균소,1、2、3、4、5、10、20、40 μmol/L cDDP급1μmol/L류련사균소여1、2、3、4、5、10、20、40 μmol/L cDDP연합처리후대세포적억제솔,금식공식분석량약간적호상작용；근거분석결과채용1 μmol/L류련사균소、3μmol/L cDDP단독혹연합작용Daoy세포24 h,류식세포의검측세포조망적변화,Western blotting검측FoxM1급조망상관단백bcl-2、bax、caspase3、취선감이린산핵당취합매(PARP)표체적변화.결과 류련사균소가농도의뢰성지억제수모세포류Daoy세포적생장.금식공식분석현시류련사균소여저농도(1、2、3、4、5μmol/L) cDDP존재명현적협동작용；류식세포분석의현시류련사균소단약조、cDDP단약조세포조망솔고우대조조,이연합용약조세포조망솔고우류련사균소단약조화cDDP단약조,차이유통계학의의(P＜0.05).Western blotting결과현시,여대조조비교,류련사균소단약조화연합용약조FoxM1단백표체명현하강,차이유통계학의의(P＜0.05)；여류련사균소단약조、cDDP단약조비교,연합용약조세포bcl-2단백표체명현하강,이bax、caspase3、PARP단백표체명현승고.결론 류련사균소화cDDP재저농도하구유명현적협동항수모세포류작용,기궤제가능여류련사균소억제FoxM1표체후증강료cDDP유도조망적능력유관.
Objective To investigate the effects of FoxM 1 inhibition by thiostrepton on CDDP chemosensitivity in medulloblastoma Daoy cells.Methods Medulloblastoma Daoy cells were cultured in vitro; the inhibitory rates of Daoy cells after the treatments of 1,1.25,1.5,1.75,2,3,5 μmol/L thiostrepton (thiostrepton treatment groups),or 1,2,3,4,5,10,20 and 40 μmol/L CDDP (CDDP treatment groups) or 1 μmol/L thiostrepton combined with 1,2,3,4,5,10,20 and 40 μmol/L cDDP (combination treatment groups) were evaluated by CCK8 assay,and the interaction between thiostrepton and cDDP was analyzed by means of Jin's formulatin.According to the analysis results,Daoy cells were treated with thiostrepton (1 μmol/L) and CDDP (3 μmol/L) combination or either drug alone for 24 h,and then,flow cytometry was used to observe the changes of apoptosis; Western blotting was employed to detect the expressions of FoxM1 and apoptosis-relating proteins of bcl-2,bax,caspase 3 and poly-ADP-ribose polymerase (PARP).Results Thiostrepton and cDDP used alone could inhibit the proliferation of Daoy cells,and there was simple addition or synergistic effect after combined treatment (1,2,3,4 and 5 μmol/L cDDP and thiostrepton).Flow cytometric analysis showed that the apoptosis rate of thiostrepton treatment groups and CDDP treatment groups was significantly higher than that of control groups,while that of combination treatment groups was significantly higher than that of thiostrepton treatment groups and CDDP treatment groups (P＜0.05).Western blotting indicated that the FoxMl expression level in the combination treatment groups and thiostrepton treatment groups was significantly lower than that in the control group (P＜0.05); as compared with that in the thiostrepton treatment groups and CDDP treatment groups,the bcl-2 protein level was significantly lower in the combination treatment groups,while Bax,caspase 3 and PARP protein expressions were significantly increased (P＜0.05).Conclusion Thiostrepton acts synergistically with cDDP in low concentration,and its mechanism may involve in inhibiting FoxM 1 expression,and then,enhancing cDDP-induced apoptosis.